( itor recently introduced a new proofrea...)Comparing Fidelity and Performance of Proofreading PCR Enzymes

itor recently introduced a new proofreading PCR enzyme, called PLATINUM Pfx DNA polymerase. Contrary to its trade name, Pfx was derived from the Pyrococcus sp. strain KOD1 (P. sp. KOD) rather than Pyrococcus furiosus (Pfu), the source of Pfu DNA polymerase. KOD DNA polymerase was first described by Tagaki, et al.⁶, and commercialized for PCR by Toyobo. Phylogenetic analyses have shown that, although reportedly from a Pyrococcus isolate,⁶ KOD DNA polymerase is more closely related to enzymes from Thermococcus species (e.g., Vent, 9N-7) than Pyrococcus species (Pfu, Deep Vent).⁷ According to its manufacturer, the Pfx version of KOD DNA polymerase possesses hot start capability provided by neutralizing polymerase antibodies.

The manufacturers claimed that Pfx DNA polymerase exhibits greater accuracy than Pfu DNA polymerase⁸ (authors cite Tagaki, et al.⁶). Pfx was also reported to provide comparable sensitivity and higher amplification specificity compared to PfuTurbo DNA polymerase.⁸ In this report, we present the results of direct fidelity and PCR performance comparisons between PfuTurbo DNA polymerase and Pfx DNA polymerase. PCR performance is assessed with respect to product yield, sensitivity, specificity, and target-length capability.

PCR Fidelity Comparisons

PCR enzyme fidelity has been measured using a number of different methods, including DGGE analyses³ and monitoring phenotypic changes in mutational target genes (lacI¹ and p53 ²). These studies showed that Pfu DNA polymerase exhibits the lowest intrinsic error rate of any commercial thermostable DNA polymerase. For example, using a PCR mutation assay based upon the lacIOlacZa target gene,¹ Pfu DNA polymerase exhibited an average error rate 2-fold lower than that of Vent and Deep Vent DNA polymerases,¹ 3- to 6-f
'"/>

Source:

Page: All 1 2 3 4 5 6 7 8
Related biology technology :

1. prostar RT-PCR Systems for Robust High-Fidelity RNA Amplification
2. pfuturbo DNA Polymerase: A High-Performance, High-Fidelity Enzyme Ideal for PCR Cloning
3. High-Fidelity PCR with a Novel Polymerase Mixture
4. Unique Enhanced DNA Polymerase Delivers High Fidelity and Great PCR Performance
5. Expand High FidelityPLUS PCR System
6. Expand High Fidelity PCR System
7. High Fidelity PCR Master
8. Challenge the Performance of Your Hot-Start PCRs with FastStart Taq DNA Polymerase and the Novel FastStart High Fidelity PCR System
9. Combine High Yield, Great Accuracy and the Prevention of Carry-over Contamination by Using the Novel Expand High FidelityPLUS PCR System
10. Novel PCR Enhancing Factor Improves Performance of Pfu DNA Polymerase
11. PCR Performance Comparisons Between pfuturbo and Taq DNA Polymerases